| Objectives Observe the expression of molecular chaperone glucose-regulated protein 78(GRP78),response element binding 6(AFF6),and C/EBP-homologous protein(CHOP),PUMA in the kidney of rats with Diabetic kidney disease(DKD).To investigate the effect of Shenshao oral liquid on the ATF6-CHOP-PUMA signal pathway and the early application of protection against apoptosis induced by renal endoplasmic reticulum stress.Methods 1 Construction of Diabetic Diabetic Kidney Disease Rat Model 55 clean-class male 8-week-old Wistar rats were randomly divided into two parts:Control group(n=11 rats)and experimental group(n=44).Pre-treatment:The control group was given normal feed,and the experimental group was given high-fat feed.After 4 weeks of feeding,control rats(n=11)were injected intraperitoneally with 25 mg/kg citrate buffer daily;The rats in the control group received intraperitoneal injection of Streptozotocin(STZ)25 mg/kg(n=44)daily.The purpose is to make type 2 diabetic rat model,after injection,fasting plasma glucose(FBG)was collected from rat tail vein blood at 3d,7d,and 14 d.After three consecutive measurements,fasting blood glucose levels greater than or equal to 16.7 mmol/L(300 mg/dl)was successfully modeled for type 2 diabetic rats.After 4 weeks of continuous feeding,24-hour urinary protein(24hUA)>30mg/24 h was successfully constructed for the DKD rat model.The experimental group was randomly divided into 4 groups:(1)Model group(DKD group,n=11):Daily irrigated with an equal volume of citrate buffer;(2)Low dose of Shenshao oral Liquid(LSS group,n=11):Daily irrigated Shenshao oral Liquid 4ml/kg;(3)Medium dose of Senshao oral Liquid group(MSS group,n=11): irrigated Shenshao oral Liquid 8ml/kg;(4)High dose of Shenshao Oral Liquid(HSS group,n=11): irrigated Shenshao oral Liquid 12ml/kg.2 The changes of blood glucose,urinary protein,CR and BUN in five groups were detected.After 12 weeks of drug intervention,blood was taken from the abdominal aorta for biochemical testing,using metabolic cages to recover rats' 24 h urine,The urine protein content was measured.3 At the end of the last week,the rats were killed by cervical dislocation,and the kidney tissues were fixed with formaldehyde to make paraffin sections,followed by HE staining and Masson staining,observe the pathological features of kidney tissue in diabetic rats.4 Immunohistochemical detection The paraffin sections were used for GRP78 and CHOP immunohistochemical staining to observe the expression of markers in each group.5 Western blot Quantitative detection of GRP78,ATF6,CHOP,PUMA expression levels.6 Apoptosis of renal cells detected by TUNEL staining.The TUNEL kit was used to perform paraffin section staining of kidney tissue to detect apoptosis.Results 1 After the model was established,the control group(Con group)grew normally,the coat color was normal,and the body weight was higher than before,and the blood sugar was within the normal range;Compared with the control group,the rats in the model group and the drug group presented multiple drinking,multiple food,and multiple urine phenomenon.The coat color was coarse and dull,and gradually appeared apathetic,and the body weight decreased compared with the control group;Taking different doses of Shenqi Oral Liquid,the blood glucose level was still higher,which was significantly different from that of the model group(P<0.05).2 Biochemical Indicators Testing Compared with the control group,CR,BUN and PRO in the model group(DKD group)increased significantly(P<0.05);CR,BUN,and PRO measures decreased in the drug intervention group,and the high-dose group(HSS group)decreased most significantly(P<0.05).3 Pathological Examination Compared with the control group(Con group),the diabetic model group(DKD group)had an increased glomerular volume,increased mesangial matrix,and renal interstitial fibrosis,whereas the Shenshao oral Liquid group had milder and higher symptoms.High-dose group(HSS group)has the most obvious improvement effect.4 Comparison of Apoptosis Apoptosis in the model group(DKD group)was significantly higher than that in the control group(Con group).The apoptosis of Shenqshao oral solution group was significantly lower than that of control group(Con group).The effect of medium-dose group(MSS group)was better than that of low-dose group(LSS group).However,it was weaker than high-dose group(Ss3 group).5 Immunohistochemical analysis The expression of GRP78 and CHOP in kidney tissue of the model group(DKD group)was significantly higher than that of the control group(Con group).The Shenshao oral liquid medication group had milder symptoms than the model group,and the high-dose group(HSS group)decreased most significantly.6 Western Blotting The expression of GRP78,ATF6,CHOP and PUMA in the model group(DKD group)was significantly higher than that in the control group(Con group)and the Shenshao oral solution group(P<0.05),high-dose group(HSS group)expression decreased significantly(P<0.05).Conclusions 1 Factors such as GRP78,ATF6,CHOP,PUMA are involved in the occurrence and development of apoptosis in diabetic endometrium(DKD)endoplasmic reticulum stress.2 Shenshao oral Liquid may have protective effects on diabetic nephropathy by inhibiting the activation of ATF6-CHOP-PUMA signaling pathway to reduce apoptosis.3 Through comparison,it was found that different doses of Shenshao oral Liquid had protective effects on the damage caused by renal endoplasmic reticulum stress in diabetic rats,and high-dose Shenshao oral Liquid had a more obvious effect. |
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